Objective:To develop a genotyping method for the Junior blood type and report on a rare blood type with Jr(a-).Methods:Healthy O-type RhD+ volunteer donors of the Shenzhen Blood Center from January to May 2021 ( n=1 568) and a pedigree with difficult cross-matching ( n=3) were selected as the study subjects. Serological methods were used for proband′s blood type identification, unexpected antibody identification, and antibody titer determination. Polymerase chain reaction-sequence specific primer (PCR-SSP) method was used for typing the proband′s RHD gene. ABCG2 gene coding region sequencing and a PCR-SSP genotyping method were established for determining the genotypes of the proband and his family members and screening of Jra antigen-negative rare blood type among the 1 568 blood donors. Results:The proband′s ABO and RhD blood types were respectively determined as B and partial D (RHDDVI.3/RHD01N.01), Junior blood type Jra antigen was negative, and plasma had contained anti-D and anti-Jra. Sequencing of the ABCG2 gene revealed that the proband′s genotype was ABGG201N.01/ABGG201N.01 [homozygous c. 376C>T (p.Gln126X) variants], which is the most common Jr(a-) blood type allele in the Asian population. Screening of the voluntary blood donors has detected no Jr(a-) rare blood type. Statistical analysis of the heterozygotes suggested that the allelic frequency for ABCG2*01N.01 (c.376T) was 0.45%, and the frequency of Jr(a-) rare blood type with this molecular background was about 0.2‰. Conclusion:A very rare case of partial DVI.3 type and Jr(a-) rare blood type has been identified. And a method for identifying the Junior blood type through sequencing the coding regions of the ABCG2 gene and PCR-SSP has been established.

Objective:A genome-wide molecular characterization of FJ21351116, a strain of G3P[8]-E2 2021 collected in Fujian, China, was performed.Methods:Whole genome sequencing of FJ21351116 was performed using a high-sensitivity group A rotavirus whole genome sequencing method. Genomic characteriza-tion of the virus was assessed by nucleic acid sequence analysis using MEGA 11.0, Geneious 9.0.2 and DNASTAR software. Neutralization epitopes of VP7 and VP4 (VP8*) were analyzed using BioEdit v. 7.0.9.0 and PyMOL v. 2.5.2.Results:In this study, FJ21351116 was shown to be a G3-P[8]-I2-R2-C2-M2-A2-N2-T2-E2-H2 genotype, and the result of phylogenetic tree showed that the VP7, VP4, VP3, and NSP2-NSP5 genes of the FJ21351116 strain were related to the equine-like DS-1-like G3P[8] genes that have been detected in Japan in recent years. VP6, VP1, VP2, and NSP1 genes are closely related to G2P[4] in most countries, especially in Singapore, suggesting that this strain was formed by genetic reassortment during the evolution of equine-like G3P[8] and G2P[4]. Evolutionary relationships between the VP7/VP4 genes of FJ21351116 and Rotarix and RotaTeq vaccines suggest that the multiple mutations in both VP7 and VP4 (VP8*) neutralizing antigenic epitopes and vaccine amino acid sites. It is hypothesized that the Rotarix and RotaTeq vaccines may be less effective against equine DS-1-like G3P[8] RVA, and the sequence differences with Rotarix are higher than those with RotaTeq.Conclusions:In this study, we found a rare case of DS-1-like G3P [8] RVA strain in China. Currently, horse-like DS-1-like G3P [8] RVA is relatively rare in China and may be poorly protected by vaccine strains, emphasizing the importance of continuous monitoring of RVA strains and the development of efficient and full-coverage RVA vaccines.

Objective:To investigate the sequence characteristics and evolutionary pattern of a strain of G1P[8] genotype group A rotvirus (RVA) SC18511073 in China and to determine the differences between SC18511073 and the antigenic epitopes of RotaTeq? and Rotarix? vaccines.Methods:RT-PCR amplification of 11 segments of SC18511073 was performed using reverse transcription-polymerase chain reaction (RT-PCR), and the typing was conducted by online RVA automatic typing tool. DNAstar5.1 and Mega11.0 software were used to analyze the homology and genetic evolution of the 11 segments.Results:The genotype constellation of SC18511073 is G1P[8]-I1-R1-C1-M1-A1-N1-T1-E2-H1, and NSP4 is the E2 genotype. The VP7 and NSP3 segments of SC18511073 had high homology with the 2018 Sichuan epidemic G1P[8]-E1 strain, and the phylogenetic tree showed that they were located in the same branch. The remaining nine gene segments all had high homology with the G9P[8]-E2 type prevalent in China and were attributed to the same evolutionary branch. SC18511073 differs from RotaTeq? and Rotarix? by a total of 5 amino acid sites on 7-1a and 7-2 of VP7, and differences in the 8-1 and 8-3 regions of VP8 * antigen epitopes. Conclusions:The 2018 RVA strain SC18511073 in China is a rare G1P[8]-E2 type, which is a new strain generated by re-assortment of VP7 and NSP3 segments during the co-infection process of G1P[8]-E1 and G9P[8]-E2 RVA strains. SC18511073 has amino acid site changes in antigenic epitopes on VP7 and VP4 with RotaTeq? and Rotarix?.

Objective: To establish a fluorescence method based on turncated aptamer for the determination of bisphenol A in water. Methods: The bisphenol A truncated aptamer containing 38 bases was selected as a recognition module, and was modified with the fluorophore 6-FAM at the 5'end. The 3'end of the complementary sequence cDNA was modified with the quencher DABCYL. The standard solutions of bisphenol A and interfering compounds were configured. The detection system was established after optimizing the number of bases in cDNA, the concentration ratio of truncated aptamer to cDNA, the incubation temperature and time, and the pH of the buffer. The specificity and recovery experiments were carried out. Results: When the complementary sequence cDNA included 9 bases, the concentration ratio of the truncated aptamer to cDNA was 1:1.5, the pH value of the buffer solution was 7.5, the cDNA was incubated at 55 ℃ for 60 minutes, in the concentration range of 10-75 pmol/L, the linear regression equation was y=2 230.7x+110 825, the correlation coefficient was 0.926. The limits of detection was 3.3 pmol/L. The difference values of fluorescence intensity between tetrabromobisphenol A, estradiol, estriol, bisphenol S and bisphenol A were obviously different, so there was no significant interference to the test result. The recovery rates were 97.8%, 98.8% and 102.3% with the spiked concentrations of 20.0, 40.0 and 60.0 pmol/L. The relative standard deviations were 4.4%, 2.1% and 2.6% (n=5), respectively. Conclusion The fluorescence method based on turncated aptamer has the advantages of easy operation, high sensitivity and specificity, which can be used for the determination of bisphenol A in water.

BACKGROUND:Reprogramming somatic cells to generate pluripotent stem cells has a wide application in biomedical research. OBJECTIVE:To analyze the effect of different molecular weight proteins in chicken egg-white extract to elevate expression of pluripotent genes Oct-3/4 and Nanog in 293T cells. METHODS:The extracts of chicken egg-white were separated into more than 3 ku and less than 3 ku ingredients to be used for co-culture with 293T cells. There were four groups, 1×105 293T cells per wel , total 500μL. In the control group, 500μL culture medium was added;in the other three groups, 500μL chicken egg-white extract, more than 3 ku and less than 3 ku ingredients were respectively added. Quantitative PCR was used to determine the relative expression levels of pluripotent genes Nanog and Oct-3/4 in 293T cells. RESULTS AND CONCLUSION:By using co-culture method, more than 3 ku ingredients have a role to increase the expression of pluripotent genes Oct-3/4 and Nanog, but less than 3 ku ingredients cannot elevate the expression of pluripotent genes. This indicates that the ingredient of chicken egg-white extract to elevate the expression of pluripotent genes is more than 3 ku proteins.

BACKGROUND:Umbilical cord as childbirth waste has wide variety of sources and can be easily obtained, without any ethical and legal restrictions. Therefore, human umbilical cord mesenchymal stem cells break al the restrictions originated from other sources of mesenchymal stem cells. OBJECTIVE:To review the application of human umbilical cord mesenchymal stem cells in cartilage diseases, neuroglioma, ischemic brain injury, lung disease, liver disease and models of myocardial infarction. METHODS:The PubMed and Wanfang databases were searched by the first author using the keywords of“human umbilical cord mesenchymal stem cells, disease models, celltherapy”in English and Chinese, respectively. Seventy-three articles were searched and final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Human umbilical cord mesenchymal stem cells have multilineage differentiation capacity similar to bone marrow mesenchymal stem cells. Compared with bone marrow mesenchymal stem cells, human umbilical cord mesenchymal stem cells have lower immunogenicity. Human umbilical cord mesenchymal stem cells show certain curative effects on cartilage disease, neuroglioma, ischemic brain injury, lung disease, liver disease and myocardial infarction, indicating that human umbilical cord mesenchymal stem cells can be used for celltransplantation to treat various diseases.

Objective To investigate the effect of the replication-deficient adenovirus carrying miR-138(Ad-miR138) on the pro-liferation of human gastric cancer cell line BGC-823 and the possible mechanism .Methods The human gastric cancer cell line BGC-823 was infected with Ad-miR138 .Then the expression level of miR-138 was measured by RT-PCR .The growth curve of BGC-823 was measured using CCK-8 method .The ability of cell invasion was measured using Transwell chambers .Results After infected with Ad-miR138 ,the expression of miR-138 in BGC-823 cells was up -regulated significantly (1 .07 ± 0 .07 vs .4 .89 ± 0 .45 ,P<0 .05) .Absorbance of the 6th day decreased significantly (0 .52 ± 0 .06 vs .0 .77 ± 0 .06 ,P<0 .05) ,and the invasion ability was de-creased obviously (32 .00 ± 11 .00 vs .56 .00 ± 12 .00 ,P<0 .05) .Conclusion miR-138 can effectively suppress the proliferation and invasion of human gastric cancer cell line BGC-823 in vitro .

BACKGROUNDThere are lots of mucus, blood, inflammatory cells and necrotic material in the pick-and-smear slides, resulting in a low detection rate. Liquid-based cytologic test (LCT) has been applied for cervical cytology diagnosis successfully and widely, however it is few reported yet for sputum cytology diagnosis at present. The aim of this study is to evaluate the clinical value of LCT in sputum examination of patients with lung cancer, and to find a novel method of early diagnosis of lung cancer.

METHODSThe cytologic findings and the diagnostic rate for lung cancer were compared between LCT and conventional pick-and-smear method.

RESULTSThere were smaller area of smear membrane, clearer background, more distinctly cytologic picture and stereoscopic fell by LCT comparing with pick-and-smear method. The diagnostic rate for small cell lung cancer by LCT was significantly higher than that by pick-and-smear method (P < 0.05). After combined detection of the two methods, the diagnostic rate for lung cancer was obviously improved (85.1%), which was remarkably higher than that by pick-and-smear method alone (P < 0.01).

CONCLUSIONSIt is operated easily for LCT to be well controlled in making smear and dyeing. LCT may be a novel technique worthy of wide use. Combination of LCT with pick-and-smear method appears to be of great value in clinical application.

BACKGROUNDTo study the diagnostic value of detection of carcinoembryonic antigen (CEA), cancer antigen 125 (CA125), cancer antigen 153 (CA153) and cancer antigen 199 (CA199) in pleural fluid samples for lung cancer.

METHODSImmunoprotein quantity of CEA, CA125, CA153 and CA199 was analyzed in pleural fluid and serum from patients with lung cancer (52 cases) and in pleural fluid from non cancerous patients (50 cases) by chemiluminescence.

RESULTSThe levels of CEA, CA125, CA153 and CA199 in pleural fluid of patients with lung cancer were significantly higher than those of non cancerous patients ( P < 0.01 or P < 0.05). In lung cancer patients, the levels of CEA, CA125, CA153 and CA199 in pleural fluid were obviously higher than those in serum ( P < 0.01 or P < 0.05). The sensitivity and the specificity of CEA+CA199 were 96.2% and 96.0%, respectively.

CONCLUSIONSDetection of CEA, CA125, CA153 and CA199 in pleural fluid might be helpful for diagnosing lung cancer, and the optimal combination for assay is CEA+CA199.

Object To show that ELSD is an excellent detector for the detection of chemical compounds devoid of chromophore, such as sarsasapogenin Methods Sarsasapogenin in crude Anemar rhena asphodeloides Bunge and its preparation was determinated by RP HPLC ELSD Results The well separated chromatographic peaks show linearity with recovery of the added sample of 100 5% in crude medicinal material and 91 38% in its preparation, r= 0 999 0 Conclusion The method was advanced, reliable, simple and can be used for quality control of crude A asphodeloides and its preparations